A Novel Terahertz Metamaterial Microfluidic Sensing Chip for Ultra-Sensitive Detection.

A terahertz metamaterial microfluidic sensing chip for ultrasensitive detection is proposed to investigate the response of substances to terahertz radiation in liquid environments and enhance the molecular fingerprinting of trace substances. The structure consists of a cover layer, a metal microstructure, a microfluidic channel, a metal reflective layer, and a buffer layer from top to bottom, respectively. The simulation results show that there are three obvious resonance absorption peaks in the range of 1.5-3.0 THz and the absorption intensities are all above 90%. Among them, the absorption intensity at M1 = 1.971 THz is 99.99%, which is close to the perfect absorption, and its refractive index sensitivity and Q-factor are 859 GHz/RIU and 23, respectively, showing excellent sensing characteristics. In addition, impedance matching and equivalent circuit theory are introduced in this paper to further analyze the physical mechanism of the sensor. Finally, we perform numerical simulations using refractive index data of normal and cancer cells, and the results show that the sensor can distinguish different types of cells well. The chip can reduce the sample pretreatment time as well as enhance the interaction between terahertz waves and matter, which can be used for early disease screening and food quality and safety detection in the future.

Exploring the mechanism of Si-Ni-San against depression by UPLC-Q-TOF-MS/MS integrated with network pharmacology: experimental research.

Background: Depression is becoming an urgent mental health problem. Si-Ni-San has been widely used to treat depression, yet its underlying pharmacological mechanism is poorly understood. Thus, we aim to explore the antidepressant mechanism of Si-Ni-San by chemical analysis and in-silico methods. Methods: Compounds in Si-Ni-San were determined by ultra-high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). Then, bioactive compounds were obtained from Traditional Chinese Medicines for Systems Pharmacology Database and Analysis Platform and SwissADME, and the potential targets of which were acquired from SwissTargetPrediction. Depression-related targets were collected from GeneCards. The intersection between compound-related targets and depression-related targets were screened out, and the overlapped targets were further performed protein-protein interaction, biological functional and pathway enrichment analysis. Finally, networks of Si-Ni-San against depression were constructed and visualized by Cytoscape. Results: One hundred nineteen compounds in Si-Ni-San were determined, of which 24 bioactive compounds were obtained. Then, 137 overlapped targets of Si-Ni-San against depression were collected. AKT1, PIK3R1, PIK3CA, mTOR, MAPK1 and MAPK8 were the key targets. Furthermore, PI3K-Akt signalling pathway, serotonergic synapse, MAPK signalling pathway and neurotrophin signalling pathway were involved in the antidepressant mechanism of Si-Ni-San. It showed that components like sinensetin, hesperetin, liquiritigenin, naringenin, quercetin, albiflorin and paeoniflorin were the mainly key active compounds for the antidepressant effect of Si-Ni-San. Conclusions: This study demonstrated the key components, key targets and potential pharmacological mechanisms of Si-Ni-San against depression. These results indicate that Si-Ni-San is a promising therapeutic approach for treatment of depression, and may provide evidence for the research and development of drugs for treating depression.

Gastrodin alleviates NTG-induced migraine-like pain via inhibiting succinate/HIF-1α/TRPM2 signaling pathway in trigeminal ganglion.

BACKGROUND: Increasing evidence highlights the involvement of metabolic disorder and calcium influx mediated by transient receptor potential channels in migraine; however, the relationship between these factors in the pathophysiology of migraine remains unknown. Gastrodin is the major component of the traditional Chinese medicine Tianma, which is extensively used in migraine therapy. PURPOSE: Our work aimed to explore the analgesic action of gastrodin and its regulatory mechanisms from a metabolic perspective. METHODS/RESULTS: After being treated with gastrodin, the mice were given nitroglycerin (NTG) to induce migraine. Gastrodin treatment significantly raised the threshold of sensitivity in response to both mechanical and thermal stimulus evidenced by von Frey and hot plate tests, respectively, and decreased total contact numbers in orofacial operant behavioral assessment. We found that the expression of transient receptor potential melastatin 2 (TRPM2) channel was increased in the trigeminal ganglion (TG) of NTG-induced mice, resulting in a sustained Ca2+ influx to trigger migraine pain. The content of succinate, a metabolic biomarker, was elevated in blood samples of migraineurs, as well as in the serum and TG tissue from NTG-induced migraine mice. Calcium imaging assay indicated that succinate insult elevated TRPM2-mediated calcium flux signal in TG neurons. Mechanistically, accumulated succinate upregulated hypoxia inducible factor-1α (HIF-1α) expression and promoted its translocation into nucleus, where HIF-1α enhanced TRPM2 expression through transcriptional induction in TG neurons, evidenced by luciferase reporter measurement. Gastrodin treatment inhibited TRPM2 expression and TRPM2-dependent Ca2+ influx by attenuating succinate accumulation and downstream HIF-1α signaling, and thereby exhibited analgesic effect. CONCLUSION: This work revealed that succinate was a critical metabolic signaling molecule and the key mediator of migraine pain through triggering TRPM2-mediated calcium overload. Gastrodin alleviated NTG-induced migraine-like pain via inhibiting succinate/HIF-1α/TRPM2 signaling pathway in TG neurons. These findings uncovered the anti-migraine effect of gastrodin and its regulatory mechanisms from a metabolic perspective and provided a novel theoretical basis for the analgesic action of gastrodin.

A chemical derivatization-based pseudotargeted liquid chromatography-tandem mass spectrometry method for sensitive and high coverage determination of bile acids in human serum.

Global profiling of bile acids (BAs) is imperative for understand their function and disease pathogenesis. But it is still a challenging task, as the collision-induced dissociation (CID) fragment ions of unconjugated BAs showed low ion intensities to insufficient analysis. Herein, we developed a highly sensitive method for pseudotargeted profiling of BAs by chemical derivatization. In the developed method, a labeling reagent, 2-dimethylaminoethylamine (DMED), was adopted to label the carboxyl group of BAs. The results demonstrated that the detection sensitivities of unconjugated BAs were increased by 4-200 folds after DMED-labeling. Moreover, to profile other potential BAs not included in the 91 known targets, diverse survey experiments were performed on Qtrap-MS to search BAs for both precursor and fragment ion species, and retention index (RI) strategy was adopted to facilitate the identification of isomers. Finally, MRM-based LC-MS/MS method was validated for the pseudotargeted profiling of the BAs submetabolome with good linearity (r2 ≥ 0.990 for 89 known BAs) and high sensitivity (0.05-0.5 ng/mL for unconjugated BAs), covering unconjugated, glycine, taurine, sulfuric acid, glucuronic acid, and as well as those doubly-conjugated with above types. With this method, a total of 107 BAs, covering 54 BAs identified by authentic standards and 53 BAs candidates, were successfully determined in human serum of women with intrahepatic cholestasis of pregnancy (ICP). Multivariate analysis revealed deferentially expressed BAs. ICP disease altered the BAs profile with a reduced proportion of unconjugated, sulfate- and doubly-conjugated BAs and an increased proportion of glycine and taurine conjugates. Altered proportion and profile of BAs in ICP groups were gradually recovered during the ursodeoxycholic acid (UDCA) therapy. Overall, the strategy of DMED-labeling technique combined with diverse survey experiments is sufficiently sensitive and robust to comprehensively analysis of metabolic profiling of BAs in human serum.

Risk Prediction Model of Early-Onset Preeclampsia Based on Risk Factors and Routine Laboratory Indicators.

Background: Globally, 10-15% of maternal deaths are statistically attributable to preeclampsia. Compared with late-onset PE, the severity of early-onset PE remains more harmful with higher morbidity and mortality. Objective: To establish an early-onset preeclampsia prediction model by clinical characteristics, risk factors and routine laboratory indicators were investigated from pregnant women at 6 to 10 gestational weeks. Methods: The clinical characteristics, risk factors, and 38 routine laboratory indicators (6-10 weeks of gestation) including blood lipids, liver and kidney function, coagulation, blood count, and other indicators of 91 early-onset preeclampsia patients and 709 normal controls without early-onset preeclampsia from January 2010 to May 2021 in Peking University Third Hospital (PUTH) were retrospectively analyzed. A logistic regression, decision tree model, and support vector machine (SVM) model were applied for establishing prediction models, respectively. ROC curves were drawn; area under curve (AUCROC), sensitivity, and specificity were calculated and compared. Results: There were statistically significant differences in the rates of diabetes, antiphospholipid syndrome (APS), kidney disease, obstructive sleep apnea (OSAHS), primipara, history of preeclampsia, and assisted reproductive technology (ART) (p < 0.05). Among the 38 routine laboratory indicators, there were no significant differences in the levels of PLT/LYM, NEU/LYM, TT, D-Dimer, FDP, TBA, ALP, TP, ALB, GLB, UREA, Cr, P, Cystatin C, HDL-C, Apo-A1, and Lp(a) between the two groups (p > 0.05). The levels of the rest indicators were all statistically different between the two groups (p < 0.05). If only 12 risk factors of PE were analyzed with the logistic regression, decision tree model, and support vector machine (SVM), and the AUCROC were 0.78, 0.74, and 0.66, respectively, while 12 risk factors of PE and 38 routine laboratory indicators were analyzed with the logistic regression, decision tree model, and support vector machine (SVM), and the AUCROC were 0.86, 0.77, and 0.93, respectively. Conclusions: The efficacy of clinical risk factors alone in predicting early-onset preeclampsia is not high while the efficacy increased significantly when PE risk factors combined with routine laboratory indicators. The SVM model was better than logistic regression model and decision tree model in early prediction of early-onset preeclampsia incidence.

Upregulation of glycolytic enzyme PFKFB3 by deubiquitinase OTUD4 promotes cardiac fibrosis post myocardial infarction.

Metabolic dysregulations have emerged as a major mediator of cardiovascular disorders and fibrotic diseases. Metabolic reprogramming contributes a lot to cardiac fibroblast activation and cardiac fibrosis post-myocardial infarction (MI), yet the mechanism remains incompletely understood. Our work aimed to determine whether or not glycolytic reprogramming, regulated by phosphofructokinase-2/fructose-2,6-bisphosphatase 3 (PFKFB3), is a therapeutic target for alleviating post-MI cardiac fibrosis. Here, we showed that cardiac fibroblasts displayed cell energy phenotype toward augmented glycolysis in response to transforming growth factor-beta 1 (TGF-ß1), evidenced by significant extracellular acidification rate (ECAR) increase and lactate accumulation. The expression of glycolytic enzyme PFKFB3, a master activator of glycolysis, was up-regulated in TGF-ß1-treated cardiac fibroblasts and in cardiac fibroblasts of post-MI mice. Pharmacological inhibition of PFKFB3 by 3PO diminished TGF-ß1-mediated profibrotic phenotypes, attenuated cardiac fibrosis, and preserved cardiac functions in post-MI mice. Meanwhile, the genetic inhibition of PFKFB3 decreased the cardiac fibroblast activation and reversed the differentiated phenotypes in vitro and in vivo. Mechanistically, we identified deubiquitinase OTUD4 as a new binding protein of PFKFB3, and their interaction blocked PFKFB3 degradation via OTUD4-mediated deubiquitylation. Taken together, this work characterized a key role for PFKFB3 in cardiac fibroblast activation and suggested that inhibiting PFKFB3-involved glycolysis is an alternative way to alleviate post-MI cardiac fibrosis. KEY MESSAGES: PFKFB3, a master activator of glycolysis, was highly expressed in ischemic cardiac fibroblasts to enhance cardiac fibrosis The deubiquitinase OTUD4 was identified as a new binding protein of PFKFB3 TGF-ß1 blunted the ubiquitination-mediated degradation of PFKFB3 via OTUD4-mediated deubiquitylation Blockade of PFKFB3 contributed to ameliorating ischemia-induced cardiac fibrosis.

Rutin ameliorates inflammatory pain by inhibiting P2X7 receptor in mast cells

Rutin is a natural anti-inflammatory ingredient widely found in medicinal plants. Studies have shown that rutin inhibits mast cell degranulation and the release of inflammatory mediators. Mast cell P2X7 receptor mediates mast cell degranulation and serves as a therapeutic target for inflammatory pain. Herein, the aim of this study was to investigate whether the anti-inflammatory mechanism of rutin is related to the mast cell P2X7 receptor. Our results showed that rutin could inhibit [Ca2+]i elevation induced by 5 mM ATP or 30 μM BZATP in a concentration-dependent manner in mouse peritoneal mast cells. Rutin also suppressed the inward current mediated by P2X7 receptor. In vivo, rutin could significantly inhibit the mechanical hypersensitivity induced by 100 mM ATP that is associated with P2X7 receptor in mast cells. Moreover, molecular docking revealed the high affinity between rutin and the P2X7 receptor crystal structure. Collectively, this study demonstrated that rutin attenuated inflammatory pain by inhibiting the activity of P2X7 receptor in mast cells. (AU)

Rutin ameliorates inflammatory pain by inhibiting P2X7 receptor in mast cells.

Rutin is a natural anti-inflammatory ingredient widely found in medicinal plants. Studies have shown that rutin inhibits mast cell degranulation and the release of inflammatory mediators. Mast cell P2X7 receptor mediates mast cell degranulation and serves as a therapeutic target for inflammatory pain. Herein, the aim of this study was to investigate whether the anti-inflammatory mechanism of rutin is related to the mast cell P2X7 receptor. Our results showed that rutin could inhibit [Ca2+]i elevation induced by 5 mM ATP or 30 µM BZATP in a concentration-dependent manner in mouse peritoneal mast cells. Rutin also suppressed the inward current mediated by P2X7 receptor. In vivo, rutin could significantly inhibit the mechanical hypersensitivity induced by 100 mM ATP that is associated with P2X7 receptor in mast cells. Moreover, molecular docking revealed the high affinity between rutin and the P2X7 receptor crystal structure. Collectively, this study demonstrated that rutin attenuated inflammatory pain by inhibiting the activity of P2X7 receptor in mast cells.

Water Extract of Senecio scandens Buch.-Ham Ameliorates Pruritus by Inhibiting MrgprB2 Receptor.

Background: Senecio scandens Buch.-Ham (S. scandens) belongs to the Compositae family. As a Traditional Chinese medicine, S. scandens has been used in China to treat conjunctivitis, mastitis and vaginitis, it also has the function of antibacterial and relieving itching. Methods: Water extract of S. scandens (WSS) was prepared and its quality was controlled by HPLC. The antipruritic effects of WSS were evaluated by itch behavioral experiments. The oxazolone and compound 48/80 were induced to mice scratch behavior, scratch was recorded 30 min after sensitization. The relationship between the antipruritic mechanism and MrgprB2 on mast cell was studied by using mast cell-deficient Kit (W-sh) "Sash" mice and MrgprB2-/- mice. The mast cells were observed by toluidine blue staining. In vitro, the effects of WSS on MrgprB2 were studied by calcium imaging; The whole-cell patch clamp method recorded the MrgprB2 mediate voltage-dependent currents in mast cells. Results: The content of rutin (0.012%) and hyperin (0.014%) in the WSS were determined. WSS could ameliorate the pruritus induced by Oxazolone (inhibition was 41.19%, p = 0.004) and compound 48/80 (inhibition was 50.29%, p = 0.001). Meanwhile, WSS could reduce the number of mast cells in mice skin tissue with allergic contact dermatitis (ACD) (p = 0.002) or compound 48/80 (p = 0.013). In addition, WSS could inhibit the calcium influx (1 mg/mL: p = 0.001, 3 mg/mL: p < 0.0001) and the voltage-dependent currents induced by activation of MrgprB2 on mast cell. WSS also attenuated the calcium influx induced by compound 48/80 in HEK293 cells overexpressing MrgprB2/X2. Conclusion: These results showed that WSS could ameliorate pruritus by inhibiting MrgprB2 receptor on mast cells.

Predictive value of the complement factors B and H for women with gestational diabetes mellitus who are at risk of preeclampsia.

OBJECTIVES: We aimed to define the relationships between the serum concentrations of complement factors B and H in mid-pregnancy and the risk of preeclampsia (PE) in patients with gestational diabetes mellitus (GDM). STUDY DESIGN: We performed a prospective nested case-control study of 503 patients with GDM who attended Peking University Third Hospital between March 2018 and December 2018. 456 patients were followed until delivery and blood samples were collected between gestational weeks 24 and 28. Thirty patients developed PE, 12 developed gestational hypertension (GH), and 42 matched cases were selected as a control group. RESULTS: The incidence of PE was 5.96 %. The serum concentrations of triacylglycerol (TG), FB, and FH of women with GDM who developed PE (GDM-PE group) in mid-pregnancy were significantly higher than those of the GDM group [TG: 3.60 (2.94-4.63) vs 2.54 (2.14-3.01) mmol/L, p < 0.001; FB: 346 (314-378) vs 284 (263-323) mg/L, p < 0.001; FH: 417 ± 45 vs 379 ± 47 mg/L, p = 0.003]. Multivariate regression analysis showed that high serum concentrations of TG and FB in mid-pregnancy were related to the risk of patients with GDM developing PE [TG: odds ratio (OR) 2.035 (95 % confidence interval (CI) 1.032-4.013); FB: OR 1.018 (95 % CI 1.001-1.035)]. The area under the curve (AUC) of FB for the prediction of PE was 0.821 (95 % CI 0.722-0.921) and that for a combination of TG, FB, and FH was 0.857 (95 % CI 0.770-0.944). CONCLUSIONS: High serum FB concentration during mid-pregnancy is a potential predictor of the risk of PE in patients with GDM, and the combination of FB, FH, and TG increased this predictive value.